Programmable CAS9-recombinase fusion proteins and uses thereof

US11661590B2 · kind B2 · utility

Assignee

• President and Fellows of Harvard College · US

Inventors

• David R. Liu · Lexington, US
• Brian Chaikind · Oakland, US
• Jeffrey L. Bessen · Somerville, US

Key dates

• Filing date: Aug 9, 2017
• Grant date: May 30, 2023
• Expiry date: Aug 9, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y207/07
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Some aspects of this disclosure provide a fusion protein comprising a guide nucleotide sequence-programmable DNA binding protein domain (e.g., a nuclease-inactive variant of Cas9 such as dCas9), an optional linker, and a recombinase catalytic domain (e.g., a tyrosine recombinase catalytic domain or a serine recombinase catalytic domain such as a Gin recombinase catalytic domain). This fusion protein can recombine DNA sites containing a minimal recombinase core site flanked by guide RNA-specified sequences. The instant disclosure represents a step toward programmable, scarless genome editing in unmodified cells that is independent of endogenous cellular machinery or cell state.